To investigate the svnthyesis of muscle proteins during differentiation of chicken myoblast, cvtosolic and membrane fractions were used for both sodium dodecvl sulfate polvcrylamide gel eBectrophoresis and two-dimensional gel electrophoresis. An extensive cell fusion was observed in 4 day culture. In the protein pattern of the cvtosolic fraction from SDS-PAGE. several protein bands including 250 kDa and 46 kDa showed remarkable changes during culture. the protein of 46 kDa was the most prominent one ann its optical density was the highest in 5 day culture (OD = 1.30). In the membrane fraction, band of 19.8 kDa showed the highest absorbance with 0.93 OD at 12 hr after initial plating and decreased gradually thereafter to 0.23 in 5 nay culture. From the results of two-dimensional gel electrophoresis of cytosolic fraction, the 46 kDa spot was observed as ko separated forms from culture 2 nary culture, and the sixte of this spot was the largest in 5 nay culture. In the pattern of membrane protein, the extensive appearance of newiv synthesized Proteins was found in a naut culture, but no Prominent spot was observed throughout culture. From the results of the present clay, we found that, during myoblast differentiation, the most prominent proteins were bands of 46 kDa and 19.8 kDa in cvtosolic and membrane fraction, respectively, and the appearance of new proteins was initiated at 48 hr after initial plating, and the 46 kDa protein was predominant in the cytoplasm of late culture in which extensive cell fusion was observed.
Journal of the Korean Society of Food Science and Nutrition
/
v.32
no.8
/
pp.1292-1296
/
2003
The purpose of this study is to investigate the changes of physico-chemical properties of chilled plaice muscle, stored at 4$^{\circ}C$ for 0 ∼ 21 days, with different packaging methods (vacuum packaged with PVDC and aerobic packaged with HDPE). pH value in aerobic packaged plaice muscle (APPM) decreased from 6.3 to 6.09 at first 2 day storage, and then increased gradually during storage time. Although pH pattern of vacuum packaged plaice muscle (VPPM) was similar to that of APPM, change of pH value during storage time was slower and lower than APPM. VBN value in aerobic packaged one increased during storage time. Especially it increased significantly after 7 days of storage. While VBN value in VPPM increased only a little to 14 days. TBA value showed significant difference between APPM and VPPM. WHC of APPM was higher than that of VPPM after 7 days of storage. In electrophoretic pattern of myofibril of APPM stored for 14 days hydrolysis of heavy chain and tropomyosin was observed. However, in VPPM, some hydrolysis occurred only in heavy chain. SDS-PAGE analysis showed that hydrolysis of VPPM occurs later than that of APPM.
The study was intended to know any relations between the rice tolerance to oxyfluorfen and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were Chokoto, Aichiasahi, Agabyeo, IR 3941 and Tablei as the tolerant group, and Mushakdanti, Weld Pally, HP 1033, HP 857, and HP 907 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase and peroxidase from rice seedling) and changes in isoenzyme activity (malate dehydrogenase, peroxidase and esterase) as affected by oxyfluorfen treatment ($10^{-4}M$) was also studied. The results are summarized as follows. -Among 19 bands separated in seed proteins, two different rice groups selected in terms of tolerance were clustered in dissimilarity. This was based on 2 facts in that G band was not present in susceptible varieties and that less activity of H, N, O, P, Q, Rand S band was shown. -Among 4 bands separated in acid phosphatase, the presence of (band and lower activity of B band was specific for tolerant varieties. For 4 minor bands separated in peroxidase, the tolerant varieties had no activity in B band and higher activity in A, C, D bands. -Time-course study of isozymes as affected by $10^{-4}M$ oxyfluorfen showed that Chokoto, the tolerant varieties, had little activity in A band and consistently higher activities in Band C bands for malate dehydrogenase. For 5 bands separated in peroxidase, B band was not found in Chokoto while A, C, D, and E bands were consistently present. Esterase was separated into about 4 bands in which Chokoto had maintained higher activities in A, C and D bands.
To clarify the effect of anti-juvenile hormone analogue (AJH) on the larval ecdysis by feeding at early stage of the 4th instar, the total amount of protein and activity of chitinolytic enzymes in the integument of Bombyx mori were analyzed, PAGE pattern of the protein was observed and the morphological changes of integument during molting period were also observed and the morphological changes of integument during molting period were also observed by means of TEM. The total amount of protein was greatly increased in premolting, then reached maximum level just before ecdysis, and rapidly decreased after the larval ecdysis in the control, while in the AJH treatment, increased 12 hr later than the control and its maximum was only 82.6% of the control. Two specific proteins, which were presumed as the protein originated from endocuticle, also appeared 12 hr later than the control and were maintained to 132 hr after AJH treatment from the aspects of the Native- and SDS-PAGE patterns, although those of the control disappeared instantly after ecdysis. Chitinase and $\beta$-N-acetylglucosaminidase activities were also suppressed and delayed by AJH treatment. Furthermore, it was observed that the apolysis took place 12 hr later than the control but new epicuticle was not formed at least until 132 hr after AJH treatment. From these results, it is suggested that the larval molting process of silkworm develops 12 hr later than the control but new epicuticle was not formed at least until 132 hr after AJH treatment. From these results, it is suggested that the larval molting process of silkworm develops 12 hr later than the control by AJH treatment but no further processing takes place just after apolysis.
The experiment was carried out to study the profile of uterine specific protein during early pregnancy in sows and to test it's immunosuppressive activity. Uterine protein samples were obtained by flushing the uterine horn on Day 3, 6, 9, 12 and 15 of the estrous cycle and the pregnancy respectively and the protein concentration of each sample was determined. The change of uterine protein was examined by sodium dodecyl sulfate(SDS)-PAGE. The immunosuppressive activity of uterine secretory protein was investigated according to the lymphocyte blastogenesis response to mitogen. The results of this experiment are summarized as follows ; 1. The uterine protein during estrous cycle and early pregnancy was relatively constant up to Day 9, but increased on Day 12. Maxium total protein values were found on Day 15. The concentration of serum proteins were about 82-95 mg during estrous cycle, but decreased to about 70-82 mg during early pregnancy. 2. The proteins components similar electrophoretic patterns(PAGE) that were no differences (band ; a, b, c, d, e, f, g, I) on Days 3, 6 and 9 of the estrous cycle and pregnancy. But there were more 2 bands specifically on Day 12 of the pregnancy and on Day 15 of estrous cycle and showed more 4 bands on Day 15 of early pregnancy. They seemed to be acidoprotein and their average molecular weight were 38,000, 22,300 and 12,600. 3. When uterine protein were added 500$\mu\textrm{g}$/ml, there was no immunosuppresive activity on Day 3 of estrous cycle and lymphocyte blastogenesis was slightly suppressed on Day 3 of pregnancy. The immunosuppressive activity on Day 9 of estrous cycle and pregnancy appeared in 500$\mu\textrm{g}$/ml and 150$\mu\textrm{g}$/ml, respectively the uterine protein on Day 12 and 15 showed immunosuppresive activity, which at the level of 150$\mu\textrm{g}$/ml during non-pregnancy and at the level of 100 to 125$\mu\textrm{g}$/ml during early pregnancy, respectively.
Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.
An $\alpha$-amylase (EC 3.2.1.1) was purified that catalyses the production of a high level of maltose from starch without the attendant production of glucose. The enzyme was produced extracellularly by thermophilic Streptomyces sp. that was isolated from Thailand's soil. Purification was achieved by alcohol precipiation, DEAE-Cellulose, and Gel filtration chromatographies. The purified enzyme exhibited maximum activity at pH 6-7 and $60^{\circ}C$. It had a relative molecular mass of 45 kDa, as determined by SDS-PAGE. The hydrolysis products from starch had $\alpha$-anomeric forms, as determined by $^1H$-NMR. This maltose-forming $\alpha$-amylase completely hydrolyzed the soluble starch to produce a high level of maltose, representing up to 90%. It hydrolyzed maltotetrose and maltotriose to primarily produce maltose (82% and 62%, repectively) without the attendant production of glucose. The high maltose level as a final end-product from starch and maltooligosaccharides, and the unique action pattern of this enzyme, indicate an unusual maltose-forming system. After the addition of the enzyme in the bread-baking process, the bread's volume increased and kept its softness longer than when the bread had no enzyme.
Journal of the Korean Society of Food Science and Nutrition
/
v.18
no.4
/
pp.423-429
/
1989
This study was investigated to determine the amino acid content, fractionation and gel electrophoresis pattern of mungbean protein isolates(MPI), which were prepared from defatted mungbean flour(DMBF) of sunhwa-nogdu(SH) and conventional(C) mungbean varieties. MPI have particulary high content of lysine and low sulfur containing amino acids, even though amino acid pattern of mungbean flour and its protein isolate were found to be similar And, there was no significant difference In the amino acid composition between SH and C varieties. Further fractionation of MPI by solubility method and electrophoresis procedure was demonstrated that the fractions, such as albumin, globulin, glutelin and residue from MPI showed $6{\sim}10$ bands in SDS/PAGE pattern. And also the most of each protein fractions had lower molecular weight than 68,000 daltons and distributed more in the molecular weight range $43,000{\sim}68,000$ daltons.
The callus from the hypocotyl region of immature embryo of Citrus junos Sieb. was efficiently induced in the $\frac{1}{2}$ MS medium containing 45uM BA after 8 weeks culture. The callus was developed into the two callus type, embryogenic callus and nonembryogenic callus, which can be distinguished by visual examination depending on color and appearence. In vitro regeneration of callus established efficiently in the hormone-free MS medium from the embryogenic callus. In order to investigate the physiological changes depending on the developmental stage of embryo, the embryo was formed in the MS medium. The embryogenic and nonembryogenic callus, and the various stages of the somatic embryo were examimed the changes of esterase activity, and their isozyme patterns as well. The protein content and esterase activities was gradually increased on the developmental stages of embryo. Total protein pattern were different by the SDS-PAGE and were appeared strong band of 23 KD in the torpedo stage. The pattern of the esterase isozyme was exhibited a difference between embryogenic callus and nonembryogenic callus. It was appered pI 6.0, 8.0, 8.2 in the embryogenic callus. Also the new band of pI 4.75 was appeared in the cotyledon. These results suggest that the changes of esterase activities and isozyme patterns are importent factor in the differentiation and development of citrus.
We studied the effect of cadmium on chlorophylls and rubisco activation in Canavalia ensiformis L. leaves. Chlorophyll levels were reduced by 5.0 ${\mu}$M Cd. Rubisco activity at 5.0 ${\mu}$M Cd was significantly smaller than that at no treatment. Rubisco Content showed patterns of change similar to rubisco activity. These data suggest that rubisco activity was associated with an amount of rubisco protein, and that the activation and induction of rubisco is inhibited by Cd. The degree of intensity of 50 and 14.5 kD polypeptides identified as the large and small subunit of rubisco by SDS-PAGE analysis at 5.0 ${\mu}$M Cd was significantly lower than that at control, indicating Cd had a e f-fect on both subunits. Under the assumption that effects of Cd on rubisco may be r elated to rubisco activase, in addition to, its activity and content we re determined . The rubisco activase activity at 5.0 ${\mu}$M Cd was more decreased than the control. A similar change pattern was also observed in content of rubisco activase. Remarkable differences in the intensitiy of both the 45 kD and 41 kD band were found between at control and Cd-treatment. These results suggest that the change in the levels of rubisco activase leads to a subsequent alter action of rubisco levels.
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